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Updated: Jun 17, 2026
Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia
Published on: September 18, 2013
[Effect of YB-1 gene knockdown on human leukemia cell line K562/A02]
Wen-lin Xu1, Lei-lei Zhou, Qiao-yun Chen
1Department of Hematology, the Affiliated People's Hospital, Jiangsu University, Zhenjiang, Jiangsu, 212002 P.R. China.
Objective:
To investigate the potential effects of YB-1 gene knockdown on gene expression profile, cell growth and apoptosis in leukemia cell line K562/A02.
Methods:
The recombinant eukaryotic expression plasmid containing YB-1 short hairpin RNA (shRNA) or random-sequence (HK) were transfected into K562/A02 cells by lipofectamine mediation. cDNA microarray was performed to explore the alteration of gene expression profile when YB-1 gene expression was decreased. Expression of CARD8 and RHOC genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). The proliferative ability of the cells was determined by methyl thiazolyltetrazolium (MTT) assay and cell cycle analysis. Cell apoptosis was assayed by Annexin V-FITC/PI double labeled flow cytometry.
Results:
The levels of YB-1 mRNA and protein decreased dramatically in three positively transfected cells when compared with untransfected K562/A02 cells or K562/A02-HK thansfected cells. Gene expression profile was altered by transfection of YB-1 shRNA into K562/A02 cells. Among 47,000 genes on the microarray, 252 genes were detected to have changes, with 143 down-regulated and 109 up-regulated. They were functionally related to cell cycle progression, gene replication, metabolism, cell apoptosis, cell signal transduction, etc. An increase in CARD8 gene expression and a decrease in RHOC gene expression have been confirmed by RT-PCR in K562/A02-YBX13 cells. The introduction of exogenous YB-1 shRNA gene into K562/A02 cells resulted in decreased proliferation, higher G1, lower G2 and S ratio in cell cycle distribution in comparison with the control groups. Annexin V/PI detection indicated higher Annexin V+ ratio in the three positively transfected cells 24 hours after cells were treated with 0.5 micromol/L of As2O3.
Conclusion:
Down-regulation of YB-1 gene by shRNA-YB-1 can alter the gene expression profile in K562/A02 cells, leading to change of cell proliferation and apoptosis.
Insights
Knocking down the YB-1 gene in leukemia cells significantly alters gene expression, reducing cell growth and promoting apoptosis. This targeted approach impacts key cellular processes, offering potential therapeutic insights for leukemia.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Context:
- The Y-box binding protein 1 (YB-1) is implicated in various cellular processes, including proliferation and drug resistance, making it a potential target in cancer therapy.
- Leukemia cell line K562/A02 is a relevant model for studying leukemia pathogenesis and therapeutic interventions.
Purpose:
- To investigate the effects of YB-1 gene knockdown on the gene expression profile, cell proliferation, and apoptosis in the K562/A02 leukemia cell line.
- To explore the potential of targeting YB-1 as a therapeutic strategy for leukemia.
Summary:
- YB-1 gene knockdown using short hairpin RNA (shRNA) in K562/A02 cells led to significant alterations in gene expression, affecting pathways related to cell cycle, replication, metabolism, and apoptosis.
- The knockdown resulted in decreased cell proliferation, altered cell cycle distribution (increased G1, decreased G2/S phases), and enhanced apoptosis, particularly when combined with As2O3 treatment.
- Validation through RT-PCR confirmed changes in CARD8 (upregulated) and RHOC (downregulated) gene expression following YB-1 knockdown.
Impact:
- YB-1 gene knockdown influences multiple critical cellular pathways, suggesting its central role in leukemia cell survival and proliferation.
- These findings highlight the potential of YB-1 as a therapeutic target for leukemia, with implications for developing novel treatment strategies.
- The study provides a comprehensive gene expression profile alteration upon YB-1 knockdown, offering a basis for further mechanistic investigations.
